Characterization of melanocortin receptor subtype expression in murine adipose tissues and in the 3T3-L1 cell line

Bruce Boston, Roger D. Cone

Research output: Contribution to journalArticle

169 Citations (Scopus)

Abstract

It has been known for many years that adipocytes express high affinity ACTH and α-melanocyte stimulating hormone (MSH) binding sites, and that ACTH, α-MSH, and β-lipotropin are potent lipolytic hormones. We show here that the adipocyte response to the melanocortin peptides results from the expression of both the MC2 (ACTH) receptor as well as the newly discovered MC5 receptor. Using RT-PCR and Northern blot hybridization, high levels of MC2 receptor messenger RNA (mRNA) were found in all adipose tissues examined in the mouse, whereas MC5 receptor mRNA was found in a subset of these. Both receptor mRNAs were also found in the 3T3-L1 cell line but only after the cells had been induced to differentiate into adipocytes. This cell line was then used to characterize the pharmacological properties of the MC2 and MC5 receptor sites in situ. The MC2 receptor exhibits properties similar to the ACTH receptor characterized in adrenocortical cells, coupling to activation of adenylyl cyclase with an EC50 of approximately 1 nM. An MSH binding site characterized in these cells is presumably the MC5 receptor, based on the observation that this is the only other melanocortin receptor mRNA detected in these cells. The MC5 receptor in the 3T3-L1 adipocyte activated adenylyl cyclase in response to α-MSH stimulation. Interestingly, Nle4, D-Phe7-α- MSH (NDP-MSH), a commonly used synthetic α-MSH agonist, was a potent antagonist of the MC5 receptor expressed in the 3T3-L1 cell line. Although the agouti signaling peptide is a potent antagonist of NDP-MSH binding to the MC1 and MC4 melanocortin receptors, agouti was unable to block NDP-MSH binding in the 3T3-L1 adipocyte.

Original languageEnglish (US)
Pages (from-to)2043-2050
Number of pages8
JournalEndocrinology
Volume137
Issue number5
DOIs
StatePublished - 1996

Fingerprint

Melanocortin Receptors
3T3-L1 Cells
Melanocyte-Stimulating Hormones
Adipose Tissue
Receptor, Melanocortin, Type 2
Cell Line
Adipocytes
Corticotropin Receptors
Messenger RNA
Adenylyl Cyclases
Adrenocorticotropic Hormone
Binding Sites
beta-Lipotropin
Receptor, Melanocortin, Type 4
Melanocortins
Peptides
Northern Blotting
melanocortin 5 receptor
Hormones
Pharmacology

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Characterization of melanocortin receptor subtype expression in murine adipose tissues and in the 3T3-L1 cell line. / Boston, Bruce; Cone, Roger D.

In: Endocrinology, Vol. 137, No. 5, 1996, p. 2043-2050.

Research output: Contribution to journalArticle

@article{ffd1f9f25d574fbd8bfdf14ab9d4ef86,
title = "Characterization of melanocortin receptor subtype expression in murine adipose tissues and in the 3T3-L1 cell line",
abstract = "It has been known for many years that adipocytes express high affinity ACTH and α-melanocyte stimulating hormone (MSH) binding sites, and that ACTH, α-MSH, and β-lipotropin are potent lipolytic hormones. We show here that the adipocyte response to the melanocortin peptides results from the expression of both the MC2 (ACTH) receptor as well as the newly discovered MC5 receptor. Using RT-PCR and Northern blot hybridization, high levels of MC2 receptor messenger RNA (mRNA) were found in all adipose tissues examined in the mouse, whereas MC5 receptor mRNA was found in a subset of these. Both receptor mRNAs were also found in the 3T3-L1 cell line but only after the cells had been induced to differentiate into adipocytes. This cell line was then used to characterize the pharmacological properties of the MC2 and MC5 receptor sites in situ. The MC2 receptor exhibits properties similar to the ACTH receptor characterized in adrenocortical cells, coupling to activation of adenylyl cyclase with an EC50 of approximately 1 nM. An MSH binding site characterized in these cells is presumably the MC5 receptor, based on the observation that this is the only other melanocortin receptor mRNA detected in these cells. The MC5 receptor in the 3T3-L1 adipocyte activated adenylyl cyclase in response to α-MSH stimulation. Interestingly, Nle4, D-Phe7-α- MSH (NDP-MSH), a commonly used synthetic α-MSH agonist, was a potent antagonist of the MC5 receptor expressed in the 3T3-L1 cell line. Although the agouti signaling peptide is a potent antagonist of NDP-MSH binding to the MC1 and MC4 melanocortin receptors, agouti was unable to block NDP-MSH binding in the 3T3-L1 adipocyte.",
author = "Bruce Boston and Cone, {Roger D.}",
year = "1996",
doi = "10.1210/en.137.5.2043",
language = "English (US)",
volume = "137",
pages = "2043--2050",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "5",

}

TY - JOUR

T1 - Characterization of melanocortin receptor subtype expression in murine adipose tissues and in the 3T3-L1 cell line

AU - Boston, Bruce

AU - Cone, Roger D.

PY - 1996

Y1 - 1996

N2 - It has been known for many years that adipocytes express high affinity ACTH and α-melanocyte stimulating hormone (MSH) binding sites, and that ACTH, α-MSH, and β-lipotropin are potent lipolytic hormones. We show here that the adipocyte response to the melanocortin peptides results from the expression of both the MC2 (ACTH) receptor as well as the newly discovered MC5 receptor. Using RT-PCR and Northern blot hybridization, high levels of MC2 receptor messenger RNA (mRNA) were found in all adipose tissues examined in the mouse, whereas MC5 receptor mRNA was found in a subset of these. Both receptor mRNAs were also found in the 3T3-L1 cell line but only after the cells had been induced to differentiate into adipocytes. This cell line was then used to characterize the pharmacological properties of the MC2 and MC5 receptor sites in situ. The MC2 receptor exhibits properties similar to the ACTH receptor characterized in adrenocortical cells, coupling to activation of adenylyl cyclase with an EC50 of approximately 1 nM. An MSH binding site characterized in these cells is presumably the MC5 receptor, based on the observation that this is the only other melanocortin receptor mRNA detected in these cells. The MC5 receptor in the 3T3-L1 adipocyte activated adenylyl cyclase in response to α-MSH stimulation. Interestingly, Nle4, D-Phe7-α- MSH (NDP-MSH), a commonly used synthetic α-MSH agonist, was a potent antagonist of the MC5 receptor expressed in the 3T3-L1 cell line. Although the agouti signaling peptide is a potent antagonist of NDP-MSH binding to the MC1 and MC4 melanocortin receptors, agouti was unable to block NDP-MSH binding in the 3T3-L1 adipocyte.

AB - It has been known for many years that adipocytes express high affinity ACTH and α-melanocyte stimulating hormone (MSH) binding sites, and that ACTH, α-MSH, and β-lipotropin are potent lipolytic hormones. We show here that the adipocyte response to the melanocortin peptides results from the expression of both the MC2 (ACTH) receptor as well as the newly discovered MC5 receptor. Using RT-PCR and Northern blot hybridization, high levels of MC2 receptor messenger RNA (mRNA) were found in all adipose tissues examined in the mouse, whereas MC5 receptor mRNA was found in a subset of these. Both receptor mRNAs were also found in the 3T3-L1 cell line but only after the cells had been induced to differentiate into adipocytes. This cell line was then used to characterize the pharmacological properties of the MC2 and MC5 receptor sites in situ. The MC2 receptor exhibits properties similar to the ACTH receptor characterized in adrenocortical cells, coupling to activation of adenylyl cyclase with an EC50 of approximately 1 nM. An MSH binding site characterized in these cells is presumably the MC5 receptor, based on the observation that this is the only other melanocortin receptor mRNA detected in these cells. The MC5 receptor in the 3T3-L1 adipocyte activated adenylyl cyclase in response to α-MSH stimulation. Interestingly, Nle4, D-Phe7-α- MSH (NDP-MSH), a commonly used synthetic α-MSH agonist, was a potent antagonist of the MC5 receptor expressed in the 3T3-L1 cell line. Although the agouti signaling peptide is a potent antagonist of NDP-MSH binding to the MC1 and MC4 melanocortin receptors, agouti was unable to block NDP-MSH binding in the 3T3-L1 adipocyte.

UR - http://www.scopus.com/inward/record.url?scp=0030010181&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030010181&partnerID=8YFLogxK

U2 - 10.1210/en.137.5.2043

DO - 10.1210/en.137.5.2043

M3 - Article

VL - 137

SP - 2043

EP - 2050

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 5

ER -